• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제36권1호
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• pp.7-15
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• 2010

Complex human tissues harbor stem cells and precursor cells, which are responsible for tissue development or repair. Recently, dental tissues such as dental pulp, periodontal ligament (PDL), dental follicle have been identified as easily accessible sources of undifferentiated cells. These tissues contain mesenchymal stem cells that can be differentiate into bone, cartilage, fat or muscle by exposing them to specific growth conditions. In this study, the authors procured the stem cell from pulp, PDL, and dental follicle and differentiate them into osteoblast and examine the bone induction capacity. Dental pulp stem cell (DPSC), periodontal ligament stem cell (PDLSC), and dental follicle precursor cell (DFPC) were obtained from human 3rd molar and cultured. Each cell was analyzed for presence of stem cell by fluorescence activated cell sorter (FACs) against CD44, CD105 and CD34, CD45. Each stem cell was cultured, expanded and grown in an osteogenic culture medium to allow formation of a layer of extracellular bone matrix. Osteogenic pathway was checked by alizarin red staining, alkaline phosphatase (ALP) activity test and RT-PCR for ALP and osteocalcin (OCN) gene expression. According to results from FACs, mesenchymal stem cell existed in pulp, PDL, and dental follicle. As culturing with bone differentiation medium, stem cells were differentiated to osteoblast like cell. Compare with stem cell from pulp, PDL and dental follicle-originated stem cell has more osteogenic effect and it was assumed that the character of donor cell was able to affect on differential potency of stem cell. From this article, we are able to verify the pulp, PDL, and dental follicle from extracted tooth, and these can be a source of osteoblast and stem cell for tissue engineering.

• Restorative Dentistry and Endodontics
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• 제48권2호
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• pp.20.1-20.13
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• 2023

This mini-review was conducted to present an overview of the use of exosomes in regenerating the dentin-pulp complex (DPC). The PubMed and Scopus databases were searched for relevant articles published between January 1, 2013 and January 1, 2023. The findings of basic in vitro studies indicated that exosomes enhance the proliferation and migration of mesenchymal cells, as human dental pulp stem cells, via mitogen-activated protein kinases and Wingless-Int signaling pathways. In addition, they possess proangiogenic potential and contribute to neovascularization and capillary tube formation by promoting endothelial cell proliferation and migration of human umbilical vein endothelial cells. Likewise, they regulate the migration and differentiation of Schwann cells, facilitate the conversion of M1 pro-inflammatory macrophages to M2 anti-inflammatory phenotypes, and mediate immune suppression as they promote regulatory T cell conversion. Basic in vivo studies have indicated that exosomes triggered the regeneration of dentin-pulp-like tissue, and exosomes isolated under odontogenic circumstances are particularly strong inducers of tissue regeneration and stem cell differentiation. Exosomes are a promising regenerative tool for DPC in cases of small pulp exposure or for whole-pulp tissue regeneration.

• Restorative Dentistry and Endodontics
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• 제35권6호
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• pp.473-478
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• 2010

연구목적: 본 연구는 인간의 치수 및 치근단 조직에서 유리된 세포를 이용하여 비교적 최근 소개된 Bioaggregate의 생체친화성을 평가하는 데에 있다. 연구 재료 및 방법: 사람의 발거치로 부터 치수 및 치근단 조직에서 배양된 세포를 이용하였다. Bioaggregate와 white MTA를 혼합하여 세포배양판에 적용한 후 같은 수의 세포를 배양하였다. 1, 3, 그리고 7일 후 위상차현미경을 사용하여 세포의 부착과 성장을 관찰하고 cell viability test를 시행하였다. 얻어진 결과는 Student t-test및 one way ANOVA를 이용하여 분석하였다. 결과: 두가지 종류의 세포 모두 Bioaggregate와 MTA가 혼합된 배양판에서 잘 성장하였으며 Bioaggregate군에서는 inhibition zone이 관찰되지 않았다. Cell viability test에서 두 그룹간 통계적인 유의성 차이는 없었다. 결론: Bioaggreagete는 치수 및 치근단 세포에 대하여 MTA와 유사한 세포친화성을 보였다.

• Restorative Dentistry and Endodontics
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• 제34권5호
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• pp.415-423
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• 2009

$\beta$-glycerophosphate는 치수의 상아모세포 분화를 촉진하는 물질이다. Portland cement는 수중에서 장기간에 걸쳐 용해되기 때문에 $\beta$-glycerophosphate을 혼합한 Portland cement는 수산화칼슘과 함께 $\beta$-glycerophosphate를 장기간 용출하게 된다. 본 실험에서는 $\beta$-glycerophosphate을 혼합한 Portland cament에 대한 인간치수세포의 반응을 알아보았다. 인간 치수 세포에 대한 $\beta$-glycerophosphate의 효과를 알아보기 위해 다양한 농도의 $\beta$-glycerophosphate와 dexamethasone에 대한 인간 치수 세포의 ALP activity을 측정하였고 alizarin red S로 염색하여 관찰하였다. $\beta$-glycerophosphate가 다양한 농도(10 mM, 100 mM, 1 M)로 혼합된 Portland cement에 대한 인간 치수 세포의 MTS assay, ALP activity를 측정하고 SEM으로 관찰하였다. 치수세포의 석회화 정도를 관찰한 연구에서 $\beta$-glycerophosphate와 dexamethasone 단독으로 적용하였을 때 거의 효과가 없었으나 5 mM $\beta$-glycerophosphate와 100 nM dexamethasone을 혼합 적용하였을 때 가장 높은 ALP acticity를 보였다. 분화제를 첨가하거나 첨가하지 않은 모든 실험군에서 치수세포에 대한 독성은 관찰되지 않았으며 Portland cement에 10 mM $\beta$-glycerophosphate을 혼합한 시편의 ALP activity가 대조군에 비교하여 가장 많이 증가하였다. 결론적으로 $\beta$-glycerophosphate이 혼합된 Portland cement는 세포 독성이 없으며 첨가물이 없는 Portland cement에 비해 치수 분화 및 석회화를 더 많이 일으키므로 임상적으로 $\beta$-glycerophosphate을 혼합한 Portland cement 적용은 재료 하방에 더 많은 상아질을 형성시킬 것으로 추측된다.

• International Journal of Stem Cells
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• 제17권3호
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• pp.330-336
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• 2024

Mesenchymal stem cells in the dental tissue indicate a disposition for differentiation into diverse dental lineages and contain enormous potential as the important means for regenerative medicine in dentistry. Among various dental tissues, the dental pulp contains stem cells, progenitor cells and odontoblasts for maintaining dentin homeostasis. The conventional culture of stem cells holds a limit as the living tissue constitutes the three-dimensional (3D) structure. Recent development in the organoid cultures have successfully recapitulated 3D structure and advanced to the assembling of different types. In the current study, the protocol for 3D explant culture of the human dental pulp tissue has been established by adopting the organoid culture. After isolating dental pulp from human tooth, the intact tissue was placed between two layers for Matrigel with addition of the culture medium. The reticular outgrowth of pre-odontoblast layer continued for a month and the random accumulation of dentin was observed near the end. Electron microscopy showed the cellular organization and in situ development of dentin, and immunohistochemistry exhibited the expression of odontoblast and stem cell markers in the outgrowth area. Three-dimensional explant culture of human dental pulp will provide a novel platform for understanding stem cell biology inside the tooth and developing the regenerative medicine.

• 대한치과보존학회:학술대회논문집
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• 대한치과보존학회 2003년도 제120회 추계학술대회 제 5차 한ㆍ일 치과보존학회 공동학술대회
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• pp.550-550
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• 2003

IL-1${\alpha}$ and TNF-${\alpha}$ play an important role in initiating and coordinating the cellular events that make up the immune response to infection. The purpose of this study was to examine the effects of proinflammatory cytokines on mineralization and HO-1 protein expression in the human pulp cells. Human pulp cell cultures between the fifth and sixth passage were used in this study. Alkaline phosphatase and osteonectin were selected as markers for mineralization that is, odontoblast-like differentiation.(omitted)

• International Journal of Oral Biology
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• 제43권2호
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• pp.77-82
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• 2018

The mesenchymal stem cells (MSCs) that reside in dental tissues hold a great potential for future applications in regenerative dentistry. In this study, we used human dental pulp cells, isolated from the molars (DPCs), in order to establish the organoid culture. DPCs were established after growing pulp cells in an MSC expansion media (MSC-EM). DPCs were subjected to organoid growth media (OGM) in comparison with human dental pulp stem cells (DPSCs). Inside the extracellular matrix in the OGM, the DPCs and DPSCs readily formed vessel-like structures, which were not observed in the MSC-EM. Immunocytochemistry analysis and flow cytometry analysis showed the elevated expression of CD31 in the DPCs and DPSCs cultured in the OGM. These results suggest endothelial cell-prone differentiation of the DPCs and DPSCs in organoid culture condition.

• Biomaterials and Biomechanics in Bioengineering
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• 제2권2호
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• pp.61-69
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• 2015

Undifferentiated stem cells are being studied to obtain information on the therapeutic potential of isolates that are produced. Dental Pulp Stem Ccell (DPSC) may provide an abundant supply of highly proliferative, multipotent Mesenchymal Stem Cells (MSC), which are now known to be capable of regenerating a variety of human tissues including bone and other dental structures. Many factors influence DPSC quality and quantity, including the specific methods used to isolate, collect, concentrate, and store these isolates once they are removed. Ancillary factors, such as the choice of media, the selection of early versus late passage cells, and cryopreservation techniques may also influence the differentiation potential and proliferative capacity of DPSC isolates. This literature review concludes that due to the delicate nature of DPSC, more research is needed for dental researchers and clinicians to more fully explore the feasibility and potential for isolating and culturing DPSCs extracted from adult human teeth in order to provide more accurate and informed advice for this newly developing field of regenerative medicine.

• International Journal of Oral Biology
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• 제37권4호
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• pp.167-173
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• 2012

This study investigated the genes involved in the differentiation of odontoblasts derived from human dental pulp stem cells (hDPSCs). hDPSCs isolated from human tooth pulp were validated by fluorescence activated cell sorting (FACS). After odontogenic induction, hDPSCs were analyzed investigated by Alizaline red-S staining, ALP assay, ALP staining and RT-PCR. Differential display-polymerase chain reaction (DD-PCR) was performed to screen differentially expressed genes involved in the differentiation of hDPSCs. By FACS analysis, the stem cell markers CD24 and CD44 were found to be highly expressed in hDPSCs. When hDPSCs were treated with agents such as ${\beta}$-glycerophosphate (${\beta}$-GP) and ascorbic acid (AA), nodule formation was exhibited within six weeks. The ALP activity of hDPSCs was found to elevate over time, with a detectable up-regulation at 14 days after odontogenic induction. RT-PCR analysis revealed that dentin sialophosphoprotein (DSPP) and osteocalcin (OC) expression had increased in a time-dependent manner in the induction culture. Through the use of DD-PCR, several genes were differentially detected following the odontogenic induction. These results suggest that these genes may possibly be linked to a variety of cellular process during odontogenesis. Furthermore, the characterization of these regulated genes during odontogenic induction will likely provide valuable new insights into the functions of odontoblasts.

• The Korean Journal of Physiology and Pharmacology
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• 제28권5호
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• pp.423-433
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• 2024

Dental pulp stem cells (DPSCs) are a type of adult stem cell present in the dental pulp tissue. They possess a higher proliferative capacity than bone marrow mesenchymal stem cells. Their ease of collection from patients makes them well-suited for tissue engineering applications, such as tooth and nerve regeneration. Chios gum mastic (CGM), a resin extracted from the stems and leaves of Pistacia lentiscus var. Chia, has garnered attention for its potential in tissue regeneration. This study aims to confirm alterations in cell proliferation rates and induce differentiation in human DPSCs (hDPSCs) through CGM treatment, a substance known for effectively promoting odontogenic differentiation. Administration of CGM to hDPSC cells was followed by an assessment of cell survival, proliferation, and odontogenic differentiation through protein and gene analysis. The study revealed that hDPSCs exhibited low sensitivity to CGM toxicity. CGM treatment induced cell proliferation by activating cell-cycle proteins through the Wnt/β-catenin pathway. Additionally, the study demonstrated that CGM enhances alkaline phosphatase activation by upregulating the expression of collagen type I, a representative matrix protein of dentin. This activation of markers associated with odontogenic and bone differentiation ultimately facilitated the mineralization of hDPSCs. This study concludes that CGM, as a natural substance, fosters the cell cycle and cell proliferation in hDPSCs. Furthermore, it triggers the transcription of odontogenic and osteogenic markers, thereby facilitating odontogenic differentiation.